# Molecular basis of γδ T lineage specification

> **NIH NIH P01** · RESEARCH INST OF FOX CHASE CAN CTR · 2021 · $2,004,349

## Abstract

PROJECT SUMMARY/ABSTRACT
Despite the growing appreciation for the critical role played by γδ T cells in host defense and immunopathology,
the molecular events controlling their development and effector function remain poorly understood. Our program
seeks to fill this gap in knowledge using genome-wide approaches that ultimately focus on key regulatory nodes.
During the last funding cycle, we provided compelling evidence that the commitment of T cell progenitors to the
αβ and γδ T cell fates depends on differences in T cell receptor (TCR) signal strength. These signaling
differences regulate fate by proportional induction of Id3, which causes graded repression of the function of E
box DNA binding proteins (E proteins). Using genome-wide approaches to define changes in E protein-DNA
binding, we generated a number of novel insights into the control of γδ T lineage specification, which can be
distilled into 3 themes: Theme 1) E protein specificity – E protein family members exhibit distinct responses to
the TCR signals of differing intensity/duration that specify fate and play distinct roles in supporting γδ T cell
development (Proj1/3/4); Theme 2) Non-coding transcription - E protein binding is extensively associated long
non-coding RNAs (lncRNA), such as ThymoD (all Projects), Importantly, Project 4 has developed novel imaging
approaches to study lncRNA function, by visualizing lncRNA promotion of chromosome looping in real time in
live cells; and Theme 3) Human γδ development - Project 3 developed a novel human pluripotent stem cell
(PSC) based system for studying human γδ T cell development, which revealed a requirement for HEB. In our
renewal application, we will explore these themes by integrating the complementary skills of the four project
leaders. The Zúñiga-Pflücker lab, together with Michele Anderson, will focus on the specific roles of E protein
family members, and their responsiveness to Id-mediated repression, in orchestrating lineage commitment in
mouse (theme 1) and human (theme 3) differentiation models. Their observations will inform the efforts of the
Murre lab to understand the molecular basis by which E protein family members control ThymoD expression, a
lncRNA (theme 2), which orchestrates both the onset of αβ lineage commitment and the ultimate loss of αβ fate
potential upon γδ lineage commitment. The efforts of both the Zúñiga-Pflücker and Murre labs will enable the
Wiest lab (with Dietmar Kappes) to understand the role of E protein family members and non-coding transcription
(theme 2) in specifying the IL-17 producing effector fate through E protein binding sites near the Tcf7 and Zbtb7b
loci. Finally, all Projects will inform efforts of Zhuang to understand how E protein family members (theme 1) and
non-coding transcription (theme 2) control the generation and function of the stereotyped γδ TCR complexes
that drive NKγδT cell development. All projects will continue to rely on the genomic expertise of Core B to assess
the imp...

## Key facts

- **NIH application ID:** 10226992
- **Project number:** 5P01AI102853-08
- **Recipient organization:** RESEARCH INST OF FOX CHASE CAN CTR
- **Principal Investigator:** DAVID L. WIEST
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $2,004,349
- **Award type:** 5
- **Project period:** 2014-05-15 → 2024-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10226992

## Citation

> US National Institutes of Health, RePORTER application 10226992, Molecular basis of γδ T lineage specification (5P01AI102853-08). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10226992. Licensed CC0.

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